CROI 2016 Abstract eBook

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Poster Abstracts

(dpi). Control and treated animals were grouped and sacrificed with AIDS or by 84 dpi. Blood studies were performed longitudinally. A full SIV necropsy was performed, including quantitative analysis of CNS and cardiac tissues. Results: Histologic and blood chemical analyses revealed no signs of PA300 mediated pathology. pK analysis demonstrated that a biologically effective concentration of PA300 (0.7uM) was achieved at 30 mg/kg in plasma and all organs tested including brain. 7/8 (87.5%) of the untreated animals developed AIDS with 4/8 (50.0%) having SIV encephalitis. By contrast only 4/11 (36.3%) of the 30mg/kg dosed PA300 treated animals developed AIDS and 0/11 (0.0%) had SIVE. Levels of inflammatory CD14+CD16+monocytes in the PA300 Rx group were significantly lower than in untreated animals. Plasma viral loads and CD4+ T cells did not differ. PA300 treated animals had significantly lower numbers of CD68+ and CD163+macrophages in CNS cortex and CD68+ and MAC387+macrophages in the left ventricle of the heart. There were reduced numbers of SIV-RNA positive macrophages in CNS with PA300 Rx. Aortic intima media thickness and cardiac ventricular fibrosis was decreased with PA300 Rx compared to controls and were correlated with decreased numbers of CD68+ and MAC387+macrophages. Conclusions: These data demonstrate that macrophage targeted therapy with PA300 resulted in prevention of AIDS and reversal of trends for the development of SIVE and cardiovascular disease in a SIV model . These data underscore the potential use of PA300 as an adjunctive therapy with ART to target activated and infected monocyte/macrophages in HIV-infected humans. 274 IL-7 and Chemokines Trigger Intestinal Cell Homing in SIV-Infected Macaques Rosalie Ponte 1 ; Magali Rancez 2 ; Suzanne Figueiredo 3 ;Véronique Fabre-Mersseman 3 ; Jacques Dutrieux 4 ; Bénédicte Charmeteau-De-Muylder 3 ;Thomas Guilbert 5 ; Rémi Cheynier 5 ; Anne Couedel-Courteille 6 1 Rsr Inst of the McGill Univ Hlth Cntr, Montréal, QC, Canada; 2 CNRS, UMR8104, Paris, France; 3 INSERM U1016, Paris, France; 4 Univ Paris Diderot, Paris, France; 5 INSERM, Paris, France; 6 Univ Diderot Paris 7, Sorbonne Paris-Cité, Paris, France Background: SIV/HIV infection induces gut mucosa damages, where the ileum represents a major site for early viral replication. Therefore, understanding T-cell and macrophage anatomical distribution in the ileummucosa along with local chemokine/cytokine network is warranted. In SIV-infected Chinese rhesus macaque (RM), we investigated SIV- induced modifications of T-cell and macrophage distribution in the ileum lamina propria (LP) and submucosa during the first 2 weeks of infection. In addition, we assessed mucosal expression of Interleukin-7 (IL-7) and chemokines involved in cell homing to the gut. Methods: Simian gut tissues were analyzed after euthanasia at day 3, 7, 10, 14 in 9 SIV-infected RM and 2 uninfected controls. Four additional uninfected macaques were sacrificed at 0.5, 1 or 7 days following IL-7 injection (80µg/Kg). Viral DNA and RNA were quantified by qPCR. The levels of mRNA coding for IL-7 and 13 chemokines involved in lymphoid and myeloid cells homing were quantified by RT-qPCR. Confocal imaging after immunohistofluorescent staining was used to investigate immune cell counts and distribution in large ileum sections (25 mm 2 ). The evolution of peripheral T-cell subsets and monocytes was assessed in circulating blood by flow cytometry. Results: Concomitant with viral detection, IL-7 overexpression was observed by day 3 in the ileum (p<0.01). The expression of CCL2, CCL3, CCL4, CCL5, CCL19, CCL25 and CXCL10 was increased by day 10 and dropped at day 14. These local chemokine expressions correlated either to viral load or to IL-7 expression in the gut. CCL2, CCL4, CCL25, CCL28 and CXCL8 were also enhanced in the ileum of IL-7-treated uninfected macaques. The number of CD8 T-cell per tissue section reached a maximum at day 10 in the LP (p<0.01) and CD4 T-cell counts were not affected. CD4 + PM-2K + macrophages significantly increased at day 3 in the LP (p<0.05) and inversely decreased in the submucosa (p<0.05). In the blood, T-cell subsets and monocyte counts transiently decreased up to day 10. Conclusions: These observations demonstrate that IL-7 expression characterizes the ileum of acutely SIV-infected Chinese rhesus macaques. IL-7- and viral replication-driven local chemokine production leads to CD8 T-cell infiltration in the tissue as well as macrophage relocalization close to the epithelium. The rapid changes in the T and myeloid cell distribution in the ileum following SIV infection highlight the importance of early intervention to prevent mucosal pathogenesis. 275LB Apoptosis of Innate Lymphoid Cells Precedes CD4 T-Cell Death in Acute SIV Infection Joseph C. Mudd 1 ; Alexandra Ortiz 1 ; Claire Deleage 2 ; Kenta Matsuda 1 ; FanWu 1 ;Vanessa Hirsch 1 ; R. Keith Reeves 3 ; Jacob D. Estes 4 ; Jason M. Brenchley 5 1 NIAID, NIH, Bethesda, MD, USA; 2 Frederick National Laboratory, Leidos BioMed Rsr, Frederick, MD, USA; 3 Harvard Univ, Boston, MA, USA; 4 Frederick Natl Lab, Leidos Biomed Rsr, Frederick, MD, USA; 5 Frederick Natl Lab for Cancer Rsr, Frederick, MD, USA Background: Innate lymphoid cells (ILC) are a newly appreciated immune cell subset that are important in health and disease. Previous reports have shown NKp44+ type 3 ILC (ILC3) that are important sources of IL-17 and IL-22 are depleted in gut tissues in HIV and SIV infection, yet it is unclear how HIV infection affects the frequencies or function of ST2+ ILC2 that secrete IL-13 or ILC1 that secrete IFN-g. Finally, the role of microbial translocation on ILC survival and function is poorly understood. Methods: In non-human primate models of HIV infection, we assessed the frequencies of ILC subsets in gut-draining mesenteric lymph nodes (MLN) by flow cytometry and IL-17, IL-13, and IFN-g production by PMA/ionomycin treatment. To determine the role of microbial translocation on ILC frequency and function, we treated healthy uninfected rhesus macaques (RM) with DSS to induce chronic gut barrier damage. Results: Bulk CD4 T cells or IL-17+ CD4 T cells (Th17) in the MLN were not lost at early infection time points (day 10-14dpi), yet NKp44+ ILC3 in acutely infected RM (N=8) were significantly depleted (mean ILC3=0.03%) when compared to ILC3 of uninfected RM (N=10)(mean ILC3= 0.12%)(p=0.007). No significant differences in frequencies of ST2+ ILC2 or ILC1 were observed between acutely infected and uninfected RM. While caspase-3 expression of MLN CD4 T cells was unchanged, caspase-3 expression was elevated in ILC1 (mean caspase-3=12%), ILC2 (mean caspase-3=24%), and ILC3 (mean caspase-3=4%) subsets in acutely infected animals when compared to caspase-3 expression in these ILC subsets in MLN of healthy animals (p ILC1 =0.001)(p ILC2 =0.04)(p ILC3 =0.002). ILC3 in chronic SIV+MLN secreted more IL-17 (mean IL-17=30.5%) than NKp44+ ILC3 from uninfected RMs when stimulated (mean IL-17= 6.6%)(p=0.004). When healthy animals were treated with DSS to induce microbial translocation (N=4), frequencies of all ILC subsets remained unchanged yet ILC3 secreted more IL-17 (mean IL-17= 33%) than ILC3 from uninfected RM (mean IL-17=6.6%)(p=0.009). Conclusions: Depletion of NKp44+ ILC3 and elevated apoptosis in all ILC subsets in the MLN is surprising given that this precedes MLN CD4 T cell death and ILCs are not permissive to SIV infection. Microbial products alone may not contribute to ILC loss, yet may augment IL-17 production in ILC3 as this is apparent in settings of microbial translocation with and without viral replication. 276 Immune Activation Profile AssociatedWith Metabolic Syndrome in HIV-Treated Patients Christina Psomas 1 ; MehwishYounas 2 ; Christelle Reynes 3 ; Celine Fernandez 1 ;Vincent Le Moing 1 ; Claudine Barbuat 4 ; Nicolas Nagot 1 ; Pierre Portales 1 ; Jacques Reynes 1 ; Pierre Corbeau 2 1 Univ Hosp Montpellier, Montpellier, France; 2 Inst of Human Genetics, Montpellier, France; 3 Inst for Functional Genomics, Montpellier Univ, UMR5203, Montpellier, France; 4 Univ Hosp Nimes, Nimes, France Background: Immune activation in HIV-1-infected patients persists under suppressive antiretroviral treatment and may fuel comorbidities such as atherothrombosis, osteoporosis, metabolic syndrome, neurocognitive disorders, and liver steatosis. Our hypothesis was that treated patients present with distinct profiles of immune activation and that each profile is linked to a specific comorbidity. We thus explored the profile of immune activation that was associated with the presence of a metabolic syndrome. Methods: We measured by flow cytometry and ELISA the level of activation of CD4+ and CD8+ T cells, B cells, monocytes, NK cells, and endothelial cells as well as of inflammation with a total of 68 soluble and cell surface markers in 120 virologically suppressed patients and 20 healthy donors (aged ≥ 45 years). We used a hierarchical clustering analysis to classify the patients according to different markers of immune activation, and logistic regression with odds ratios (OR) and 95% CIs to measure the association between immune activation profiles and metabolic syndrome. Results: We observed evidence of inflammation and immune activation in all the cell subpopulations analysed. Patients were clustered in 5 distinct immune activation profiles. Each one of these 5 profiles could be characterized by a marker of CD8+ T cell, NK cell, monocyte, endothelial cell activation or of inflammation, respectively, and could be

Poster Abstracts

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CROI 2016